1. Volume 19, Issue 2, Pages 246-254 (April 2017)
Uroplakin 3a+ Cells Are a Distinctive Population of Epithelial Progenitors that Contribute to Airway Maintenance and Post-injury Repair 
Arjun Guha, Aditya Deshpande, Aradhya Jain, Paola Sebastiani, Wellington V. Cardoso 
Cell Reports 
Volume 19, Issue 2, Pages (April 2017)
DOI: /j.celrep
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2. Cell Reports 2017 19, 246-254DOI: (10.1016/j.celrep.2017.03.051)
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3. Figure 1
Enrichment around NEBs and Lack of Cyp2f2 Expression in Some Cells Suggest that Upk3a-Expressing Cells May Be Related to V-CCs
(A) Expression patterns of tdT (red), Scgb1a1 (green), and the pulmonary neuroendocrine cell (PNEC) marker Cgrp (white) in thick sections (300 μm) and thin sections (5 μm, right). Note airway branchpoint NEB-associated (arrows), non-branchpoint NEB-associated (circled), and solitary (arrowheads) tdT+ cells. Inset: double ISH/IHC for Upk3a mRNA, tdT protein (red), and PGP9.5 (green). Arrows indicate Upk3a mRNA+ tdT− cells. The arrowhead indicates Upk3a mRNA+ tdT+ cells.
(B) Distribution of tdT (red) and Scgb1a1 (green) at the BADJ.
(C) Quantitation of total and branchpoint NEBs associated with tdT+ cells (mean percent association with tdT+ cells ± SD, n = 3 animals).
(D) Immunostaining for tdT (red), Scgb1a1 (green), and Cyp2f2 (white). Note a pair of tdT+ cells (arrows) that express Scgb1a1 but do not express Cyp2f2; other Scgb1a1+ cells express high levels of Cyp2f2. tdT was detected by direct immunofluorescence in thick sections (also see text).
∗ p < 0.05, statistically significant (see text for details). See also Figure S1.
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4. Figure 2
Upk3a-Expressing Cells Survive Nap and Proliferate after Nap to Generate CCs and Ciliated Cells
(A) Experimental scheme to investigate the response of Upk3a-expressing cells to Nap injury.
(B) Immunostaining for tdT (red) and Scgb1a1 (green) in mock-treated (control [CTRL]) and Nap-treated lungs. Scgb1a1+ cells (green) were depleted in Nap-injected animals but not in animals injected with the vehicle alone. Note a single tdT+ Scgb1a1+ cell that survived Nap injury.
(C) Expression patterns of tdT (red), Scgb1a1 (green), and Cgrp (white) in a thick section (300 μm) from a lung 2 days after Nap. Arrows indicate NEBs associated with tdT+ cells. Arrowheads indicate solitary tdT+ cells. The circled region indicates a patch of Scgb1a1+ tdT− cells.
(D) Immunostaining for tdT (red) and Scgb1a1 (green) in Nap-treated lungs 30 days after Nap.
(E) Quantitation of frequencies of tdT+ cells in the different conditions (mean cell frequency/section ± SD). The difference between CTRL30d and Nap30d is statistically significant (see text).
(F) Quantitation of total and branchpoint NEBs associated with tdT+ cells 30 days after Nap (mean percent association with tdT+ cells ± SD, n = 3 animals).
(G) Immunostaining for tdT (red), Ki-67 (green), and Scgb1a1 (white) 7 days after Nap.
(H) Immunostaining for tdT (red), Scgb1a1 (green), and β-tubulin IV (white) 30 days after Nap. β-tubulin IV+ cells are indicated by arrows.
(I) Quantitation of cell types (tdT+ cells, n = 3 animals) 30 days after Nap.
A, airway; ∗ p < 0.05, statistically significant (see text for details). See also Figure S2.
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5. Figure 3
Upk3a-Expressing Cells Contribute to Airway Maintenance Long-Term and Generate Both CCs and Ciliated Cells
(A) Experimental scheme to investigate the contribution of Upk3a-expressing cells to airway maintenance.
(B) Quantitation of frequencies of tdT+ cells at different time points (mean cell frequency/section ± SD; control, 3 months, and 6 months, n = 4 animals; 9 months, n = 3 animals).
(C–E) Immunostaining for tdT (red) and Ki-67 (green) at 3 months (C), 6 months (D), and 9 months (E) after induction.
(F) Expression patterns of tdT (red), Scgb1a1 (green), and CGRP (white) in a thick section (300 μm) from a lung 9 months after induction. Note airway branchpoint NEB-associated (arrows), non-branchpoint NEB-associated (circled), and solitary (arrowheads) tdT+ cells.
(G) Quantitation of total and branchpoint NEBs associated with tdT+ cells 9 months after induction (mean percent association with tdT+ cells ± SD, n = 2 animals).
(H) Immunostaining for tdT (red) Scgb1a1 (green), and β-tubulin IV (white) 9 months after induction. The arrow denotes a tdT+ Scgb1a1+ cell, the arrowhead denotes a tdT+ β-tubulin IV+ cell.
(I) Quantitation of cell types (tdT+ cells) (control, 3 months and 6 months, n = 4 animals; 9 months, n = 3 animals).
See also Figure S2.
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6. Figure 4
Upk3a-Expressing Cells Contribute to Alveolar Repair after Bleo Injury
(A) Experimental scheme used to investigate contribution of Upk3a-expressing cells to alveolar repair after Bleo injury.
(B) Immunostaining for tdT (red) and Scgb1a1 (green) 25 days after Bleo.
(C and D) Immunostaining for tdT (red), Pdpn (AT1, green), and Sftpc (AT2, white) 25 days after Bleo showing tdT+ cells in the parenchyma that are positive or negative for Pdpn/Sftpc. Arrows indicate AT1 (Pdpn+) cells, short arrows indicate AT2 (Sftpc+) cells, and arrowheads indicate tdT+ Pdpn− Sftpc− cells.
(E) Immunostaining for tdT (red), Pdpn (AT1, green), and Sftpc (AT2, white) in the lung parenchyma 4 months after Bleo. Arrows indicate AT1 (Pdpn+) cells, and short arrows indicate AT2 (Sftpc+) cells.
See also Figure S2.
Cell Reports  , DOI: ( /j.celrep )
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